HLA-High Throughput Sequencing

Birgitte B. Simen, R&D Molecular Biology, Roche, USA

Learning Objectives:

Upon the completion of this presentation the participants will be able to:

1. Explain why setting phase is important for HLA typing
2. Contrast population-based and high throughput clonal sequencing
3. Recognize the utility of full-length clonal exon sequencing for genotyping

The high degree of polymorphism at human leukocyte antigen (HLA) class I and class II loci makes high-resolution HLA typing challenging. Current typing methods, including population-based (Sanger) sequencing, yield ambiguous typing results because of incomplete genomic coverage and inability to set phase for HLA allele determination. The long sequencing reads from Roche 454 Life Sciences’ next generation sequencing system coupled with Conexio ATF software provides very high resolution HLA genotyping. High-throughput genotyping can be achieved by use of primers with multiplex identifier (MID) tags to allow pooling of the amplicons generated from different samples prior to sequencing.Data from both the high throughput GS FLX System and the benchtop GS Junior System will be presented.